BIOTECON Diagnostics DNA / RNA Extraction DNA / RNA Extraction – Key to Successful Analysis

StarPrep Kits ShortPrep Kits Sample Preparation Kits Magnetic Preparation Kits

Fast and easy lysis by mixing samples with buffer High purity DNA/RNA clean-up, manual column based or automated magnetic bead based

Reliable analysis of food and feed samples with real-time extraction kits have been optimized in combination with PCR require appropriate sample preparation. Optimized our foodproof® real-time PCR Detection, Identification and and validated methods for the extraction of nucleic acids Quantification Kits. The isolated DNA/RNA can be used from any kind of biological material are of high importance. directly in all our real-time PCR assays. The processed sample must be free of contaminants, especially inhibitors, while retaining even small amounts Direct lysis, Column based or Automated of the target’s genetic information. The extraction method The foodproof® ShortPrep, StarPrep, Sample Preparation should be as fast and convenient as possible without and Magnetic Preparation Kits assist you in efficiently compromising safety or reproducibility. working with all types of sample matrices, including difficult complex samples like e.g. cheeses, spices or cacao. Optimized Combination of Extraction Kit and PCR Assay BIOTECON Diagnostics has developed a broad range of Succeed the challenges of complex matrices, PCR fast, safe and easy-to-use DNA and RNA extraction kits inhibitors and hidden targets with fast, straight-forward for any food, beverage or environmental sample. Our easy-to-use solutions. DNA / RNA Extraction – Key to Successful Analysis

Table 1: Comparison of foodproof® extraction kits for food and beverage relevant parameters

Rapid Highly purified Target Bead based/ Bulk Prefilled Column based Automated Sample Magnetic Gram-negative bacteria StarPrep One* ShortPrep I Preparation I Preparation I, V

Sample Magnetic Gram-positive bacteria StarPrep Two* ShortPrep II Preparation II Preparation II

Enterobacteriaceae, Magnetic StarPrep One - - Cronobacter Preparation IV

Vibrio, STEC, StarPrep Three - - - Beer spoilage bacteria

Dekkera, S. cerevisiae var. StarPrep Two StarPrep Four - - diastaticus

Yeast & Mold StarPrep Two - - -

Sample Magnetic GMOs, Animal ID, Allergens - - Preparation III Preparation III

Sample Viruses - - - Preparation IV

* High-throughput protocol available. See 8-strip DNA extraction protocols. StarPrep Kits: Versatile

WORKFLOW e.g. StarPrep One Enjoy convenient and flexible DNA StarPrep can be used for rapid DNA extraction: the foodproof® StarPrep extraction from enrichment cultures, Kits are designed for quick and direct and rinse samples. The reagents ADD SAMPLE easy extraction of DNA from food contain no hazardous chemicals or CENTRIFUGE 5 min DISCARD SUPERNATANT and beverage samples by heat- compounds, unlike other sample induced lysis. StarPrep is a cost- preparation methods. The entire DNA effective, versatile solution for nearly preparation can be performed in a ADD LYSIS BUFFER all kind of foods, with optimized lysis single tube, minimizing handling steps, reagents and protocols for different reducing risk of cross-contamination applications. and exposure to bio-hazardous material. StarPrep One Kit Bulk solution for fast and flexible DNA extraction MIX Table 2: Overview of foodproof® StarPrep Kits

foodproof® StarPrep One foodproof® StarPrep Two foodproof® StarPrep Three foodproof® StarPrep Four

Gram-positive and other INCUBATE bacteria, e.g. Listeria, Vibrio, Dekkera, 10 min Gram-negative bacteria, Targets Campylobacter, E. coli STEC, S. cerevisiae var. e.g. Salmonella O157, Alicyclobacillus; Beer spoilage bacteria diastaticus Yeast and Mold Suitable for a wide range of food, animal feed and MIX Matrices Enrichment cultures, beverage samples environmental samples Mechanical disruption and Mechanical disruption and Principle Heat-induced lysis Heat-induced lysis heat-induced lysis heat-induced lysis CENTRIFUGE 1 min Isolations 100 / 500 96 96 96

Additional Resuspension Reagent, - - - Products Washing Solution SUPERNATANT MicroVal, AOAC, FOR DETECTION Approvals AOAC, NordVal and DAFF - - NordVal and DAFF ShortPrep Kits: Ready-to-Use

WORKFLOW The fast and easy DNA extraction method for your enriched e.g. ShortPrep I samples: the foodproof® ShortPrep Kits are designed for quick and easy DNA extraction of bacterial DNA to be used directly for PCR. Simply add enriched sample to pre-aliquoted buffer, and in less than 30 minutes, samples from bacterial enrichment cultures are ready for PCR. The entire extraction ADD SAMPLE is performed in a single tube, minimizing handling steps and to prefilled lysis buffer reducing risk of cross-contamination. The lysis buffers are free of dangerous reagents. The kits are validated and certified by internationally MIX recognized institutions such as AOAC, NordVal and DAFF, and can be used for a variety of foods, like raw materials or processed food, environmental samples and pet foods. ShortPrep I Kit Prefilled tubes for fast DNA extraction INCUBATE in less than 30 minutes 10 min Table 3: Overview of foodproof® ShortPrep Kits

foodproof® ShortPrep I foodproof® ShortPrep II MIX Gram-positive and other bacteria, e.g. Listeria, Targets Gram-negative bacteria, e.g. Salmonella Campylobacter and Alicyclobacillus

Matrices* Suitable for a wide range of food, animal feed and environmental samples CENTRIFUGE Principle Heat-induced lysis Mechanical disruption and heat-induced lysis 1 min

Convenience 96 individual ready-to-use reaction mixes for maximum efficiency and easy DNA extraction

Approvals AOAC, NordVal and DAFF SUPERNATANT FOR DETECTION * DNA extracts from some raw materials of plant origin (e.g. spices) and from some beverage samples may still contain substances that interfere with PCR amplification or with fluorescent signal detection. For such materials, preparation and purification with the BIOTECON Diagnostics’ foodproof Sample Preparation Kit I and II are recommended. Sample Preparation Kits: High Quality Purification

WORKFLOW e.g. Sample Preparation III Kit* Get rid of unwanted PCR contaminants and inhibitors: the foodproof® Sample Preparation Kits are designed to isolate and purify DNA or RNA even from challenging ADD EXTRACTION BUFFER matrices containing high amounts of PCR inhibitors. Kit to sample reagents and consumables are optimized to obtain the best results for each parameter. Protocols are easy to Sample Preparation III Kit MIX follow and implement in daily routine use. The quality of the INCUBATE 30 min Column-based DNA obtained with the kits is suitable for any application extraction for CENTRIFUGE 10 min using real-time PCR. highly purified DNA

TRANSFER SUPERNATANT

ADD BINDING BUFFER & PROTEINASE K Table 4: Overview of foodproof® Sample Preparation Kits INCUBATE 10 min ADD ISOPROPANOL foodproof® Sample foodproof® Sample foodproof® Sample foodproof® Sample Preparation I Kit Preparation II Kit Preparation III Kit Preparation IV Kit TRANSFER SUPERNATANT to filter column Gram-positive and other Genetically modified CENTRIFUGE 1 min Gram-negative bacteria, bacteria, e.g. Listeria, organisms (GMOs), Viruses Targets DISCARD FLOW THROUGH e.g. Salmonella Campylobacter and animal identification (e.g. Norovirus) REPEAT ONCE Alicyclobacillus allergens

ADD WASH BUFFER Matrices Suitable for a wide range of food, beverages, animal feed and environmental samples CENTRIFUGE 1 min The RNA is bound to a DISCARD FLOW THROUGH The DNA is bound to a glass fiber filter, washed thoroughly and the pure DNA is eluted Principle glass fiber filter, washed REPEAT ONCE from the columns and eluted

ADD ELUTION BUFFER Isolation of highly purified DNA, virtually free of other cellular components and PCR inhibitors CENTRIFUGE 1 min Isolation of viral RNA, Features Suitable for matrices containing high amount of PCR inhibitors FLOW THROUGH process control included FOR DETECTION Sample Preparation Kit II and Sample Preparation Kit III include a lysozym and Proteinase K digestion step, respectively, for maximum performance * Workflow for GMOs & allergens; Animal ID slightly different protocol Sample Preparation Kits: High Quality Purification Magnetic Preparation Kits: Automated Purification

Automation Systems BIOTECON Diagnostics offers a variety of solutions The KingFisherTM Flex is a semi- for automated high-throughput molecular testing for automated system that allows fast and pathogens, GMOs, animal identification and allergens. convenient DNA extraction. The high The foodproof® Magnetic Preparation Kits are based on throughput system processes up to 96 magnetic bead technology, providing ultra-pure, ready- samples simultaneously; fewer samples to-use genomic DNA of bacterial, plant or animal origin may also be processed. for enhanced performance in sensitive real-time PCR Magnetic Preparation Kit I applications. The process proceeds with DNA being bound Bead-based DNA to magnetic beads and washed thoroughly through multiple extraction for steps. In the final step, highly purified DNA is eluted from automation the beads and is ready for use.

Table 5: Overview of foodproof® Magnetic Preparation Kits KingFisherTM Flex

® ® ® ® ® foodproof MPK I foodproof MPK II foodproof MPK III foodproof MPK IV foodproof MPK V The Janus IntegratorTM allows for full automation of processing samples Gram-negative GMOs, Entero-bacteriaceae, Gram-positive Targets bacteria, e.g. animal identification, Cronobacter and Salmonella spp. through DNA extraction and PCR setup. bacteria, e.g. Listeria Salmonella spp. allergens Salmonella spp. The user then need only load the final samples in the PCR cycler. Matrices food and feed food and feed food and feed infant formula food

Approvals AOAC and MicroVal - - MicroVal -

Additional Consumable Pack Consumable Pack Consumable Pack Consumable Pack included Products (960 isolations) (960 isolations) (480 isolations) (480 isolations) foodproof® foodproof® Instruments RoboPrep+ HT, KingFisherTM Flex KingFisherTM Flex KingFisherTM Flex RoboPrep+ HT KingFisherTM Flex foodproof® RoboPrep+: The KingFisherTM Flex + optional Janus IntegratorTM Reagent D: Elimination of DNA from dead cells

PRINCIPLE Matrices like powdered infant formula or Elimination Procedure 1. Reagent D Treatment processed seafood may contain a high Reagent D contains light sensitive background of dead (inactive) target substances that selectively penetrate cells, which may cause false-positive cell membranes of dead cells. Exposure results or distort quantitative PCR to visible light leads to covalent binding

live cells dead cells assays. Reagent D is highly selective of Reagent D to DNA and prevents the in penetrating only membranes of masked DNA from being amplified in dead cells and can be used for live/ the PCR. Reagent D has been validated dead differentiation to avoid false- in combination with the foodproof® positive results. StarPrep Kits. Reagent D Kit 2. After Extraction Processing takes only a few minutes For elimination of false-positive results prior to performing the DNA extraction. from dead cells to avoid false-positive results

foodproof® D-Light light dose with high power LEDs and 3. Real-time PCR Run As a validated source of light for proper optical alignment guarantees DNA of live cells DNA of dead cells the photo-activation of Reagent D, reproducible results. The workflow is the stand-alone CE/FCC certified simple and safe: no cooling needed, as ® amplification amplification blocked foodproof D-Light is a compact no heat will be developed. benchtop solution that fits in every lab. The combination of the foodproof® Up to 96 samples can be processed D-Light and Reagent D, foodproof® without Reagent D with Reagent D simultaneously. SBS format allows use DNA extraction kits and subsequent

Intensity Intensity of different adapters and microplates. real-time PCR kits and all assay- life cells A pre-programmed incubation starts required laboratory equipment live + dead dead cells cells automatically after insertion of the represents a true “One-Stop-Shop” Cycles Cycles samples. A constant and uniform solution for the customer. Reagent D: Elimination of DNA from dead cells 8-Strip DNA Extraction Protocols

WORKFLOW For efficient manual handling of large sample numbers, High-Throughput Salmonella Protocol e.g. StarPrep One BIOTECON Diagnostics has developed time-saving The foodproof® StarPrep One Kit includes a high-throughput protocols for DNA extraction from Salmonella and Listeria method, which is described in the manual. Necessary enrichment cultures. instruments and consumables (please see table 6) can be ADD SAMPLE The manual high-throughput DNA extraction protocols utilize purchased from BIOTECON Diagnostics. CENTRIFUGE 10 min a validated optimized combination of consumables and DISCARD SUPERNATANT instruments, including a multichannel pipette. The 8-channel High-Throughput Listeria Protocol pipette reduces the number of handling steps, and the The foodproof® StarPrep Two 8-Strip Kit already contains ADD DILUTED 8-strip format is adaptable to the number of samples. The all necessary reagents and consumables. The ready-to use LYSIS BUFFER detailed manuals make it easy to follow the protocols. The 8-strip tubes are prefilled with beads. These tubes and the MIX obtained DNA can be used directly in any PCR application. lysis buffer are included in the kit. to resuspend pellet

Table 6: Overview of additional equipment INCUBATE 10 min Instrument StarPrep One StarPrep Two

Magnetic Stirrer X -

Heating Block X X CENTRIFUGE Centrifuge X X 5 min

Mechanical Cell - X Disruption Multichannel X X Pipette SUPERNATANT Consumables X included FOR DETECTION Basic Lab Instruments

BIOTECON Diagnostics provides its customers a “one- nucleic acid extraction and real-time PCR kits. This select stop shop” solution. We proudly offer instruments and equipment has been thoroughly tested by our team to laboratory equipment necessary to perform our high quality guarantee quality performance.

Disruptor Genie® Micro Star 17 microcentrifuge Heating-Thermomixer MHR 13 Ideal for cell disruption/homo- Powerful (17,000 ×g), safe, compact, All-in-one: heating, cooling, mixing, genization of bacteria and pellet versatile, convenient and ventilated timer with one single unit resuspensions centrifuge 30 different exchange blocks Hands-free automatically timed Easy operation, intuitive controls, available (e.g. for the use of micro disruption of up to 12 microtubes bright and easy to read displays test tubes, test tubes, centrifugal Patented multi-directional action Designed to accelerate the routine tubes, cuvettes or PCR plates, agitates and vortexes at high-speed sample preparation process microtiter or deep well plates) producing greater yields in less time Special ‘click’ biocontainment rotor lid State-of-the-art touch panel for Suitable for use in cold rooms or in keeps the user and the samples safe better handling and cleaning incubators Supplied with 24-place rotor for Possible to program up to 9 Can be also used as vortexer 1.5/2.0 ml tubes and aerosol-tight complete programs with up to 30 rotor lid steps Basic Lab Instruments

Table 7: Basic laboratory equipment required for performing nucleic acid extraction kits Magnetic Heating Reagent D & RNA/DNA Extraction Kit Disruptor Vortex Centrifuge Stirrer Block D-Light

foodproof® StarPrep One Kit X 1.5 ml Block - EbC* X ≥13,000 xg

foodproof® StarPrep Two Kit X 1.5/2 ml Block X Yeast & Mold X ≥13,000 xg

foodproof® StarPrep Three Kit X 1.5 ml Block - Vibrio X ≥13,000 xg

foodproof® StarPrep Four Kit - 1.5 ml Block X - X ≥13,000 xg

foodproof® ShortPrep I Kit - 1.5 ml Block - - X ≥8,000 xg

foodproof® ShortPrep II Kit - 1.5 ml Block X - X ≥13,000 xg

foodproof® ShortPrep III Kit - 1.5 ml Block - - X ≥15,000 xg

foodproof® Sample Preparation Kit I - 1.5 ml Block - - X ≥13,000 xg

foodproof® Sample Preparation Kit II - 1.5 ml Block - - X ≥13,000 xg

foodproof® Sample Preparation Kit III - 2 ml Block - - X ≥13,000 xg

foodproof® Sample Preparation Kit IV - - - - X ≥20,000 xg

foodproof® Magnetic Preparation Kit I ------

foodproof® Magnetic Preparation Kit II - - - - - ≥2,250 xg**

2 ml Block, foodproof® Magnetic Preparation Kit III - - - X - mix

foodproof® Magnetic Preparation Kit IV - - - EbC* - -

foodproof® Magnetic Preparation Kit V ------

* Enterbacteriaceae plus Cronobacter ** Centrifuge for deepwell plates Product List

foodproof® & Real-time microproof® PCR Cycler DNA Extraction Kits Additionals Detection Kits S 400 01 foodproof® ShortPrep I Kit, 96 reactions A 500 02 Reagent D, 30 ml PRODUCTS & S 400 02 foodproof® ShortPrep II Kit, 96 reactions A 500 02 S Reagent D, 15 ml SERVICES S 400 03 foodproof® ShortPrep III Kit, 96 reactions A 500 12 Reagent P, 550 reactions S 400 04 foodproof® Sample Preparation Kit I, 100 reactions A 500 14 Resuspension Reagent, 35 ml foodproof® Automation DNA / RNA S 400 05 foodproof® Sample Preparation Kit II, 100 reactions Extraction Kits ® Basic Lab Instruments Service Lab S 400 06.1 foodproof Sample Preparation Kit III, 50 reactions S 400 14 foodproof® Sample Preparation Kit IV, 50 reactions D 100 03 Disruptor Genie® S 400 07 foodproof® StarPrep One Kit, 100 reactions D 110 10 Micro Star 17 microcentrifuge S 400 07 L foodproof® StarPrep One Kit, 500 reactions D 110 44 Heating-Thermomixer MHR 13 S 400 08 foodproof® StarPrep Two Kit, 96 reactions D 110 38 TH 21 Heating Block S 400 17 L foodproof® StarPrep Two 8-Strip Kit, 480 reactions D 110 39 TH 21 Exchange heating block for ® deepwell plates BIOTECON S 400 18 foodproof StarPrep Three Kit, 96 reactions ® D 110 27 Magentic Stirrer Diagnostics GmbH S 400 09 foodproof StarPrep Four Kit, 96 reactions S 400 11 foodproof® Magnetic Preparation Kit I, 96 reactions Many others on request Hermannswerder 17 14473 Potsdam S 400 11 L foodproof® Magnetic Preparation Kit I, 480 reactions Germany S 400 12 foodproof® Magnetic Preparation Kit II, 96 reactions Automation Phone: +49 (0) 331-2300-200 S 400 12 L foodproof® Magnetic Preparation Kit II, 480 reactions D 220 01 KingFisherTM Flex Fax: +49 (0) 331-2300-299 S 400 13 L foodproof® Magnetic Preparation Kit III, 480 reactions Z 600 06 KingFisherTM Flex Consumable Pack [email protected] S 400 15 L foodproof® Magnetic Preparation Kit IV, 480 reactions www.bc-diagnostics.com S 400 19 L foodproof® Magnetic Preparation Kit V, 480 reactions BR-DNAEX-EN V01/2016